6PPD and its metabolite 6PPDQ induce different developmental toxicities and phenotypes in embryonic zebrafish.

The automobile tire antioxidant N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD) and its quinone metabolite 6PPDQ have recently received much attention for their acute aquatic toxicity. The present study investigated the mechanistic developmental toxicity of 6PPD and 6PPDQ in embryonic zebrafish. Neither compound induced significant mortality but significantly decreased spontaneous embryo movement and heart rate. Both compounds induced malformations with different phenotypes; the 6PPD-exposed larvae manifested a myopia-like phenotype with a convex eyeball and fusion vessels, while the 6PPDQ-exposed embryonic zebrafish manifested enlarged intestine and blood-coagulated gut, activated neutrophils, and overexpressed enteric neurons. mRNA-Seq and quantitative real-time PCR assays showed that 6PPD- and 6PPDQ-induced distinct differential gene expression aligned with their toxic phenotype. 6PPD activated the retinoic acid metabolic gene cyp26a, but 6PPDQ activated adaptive cellular response to xenobiotics gene cyp1a. 6PPD suppressed the gene expression of the eye involved in retinoic acid metabolism, phototransduction, photoreceptor function and visual perception. In contrast, 6PPDQ perturbed genes involved in inward rectifier K+ and voltage-gated ion channels activities, K+ import across the plasma membrane, iron ion binding, and intestinal immune network for IgA production. The current study advances the present understanding the reason of why many fish species are so adversely impacted by 6PPD and 6PPDQ.

High-performance optical control of GPCR signaling by bistable animal opsins MosOpn3 and LamPP in a molecular property-dependent manner.

Optical control of G protein-coupled receptor (GPCR) signaling is a highly valuable approach for comprehensive understanding of GPCR-based physiologies and controlling them precisely. However, optogenetics for GPCR signaling is still developing and requires effective and versatile tools with performance evaluation from their molecular properties. Here, we systematically investigated performance of two bistable opsins that activate Gi/Go-type G protein (mosquito Opn3 (MosOpn3) and lamprey parapinopsin (LamPP)) in optical control in vivo using Caenorhabditis elegans. Transgenic worms expressing MosOpn3, which binds 13-cis retinal to form photopigments, in nociceptor neurons showed light-induced avoidance responses in the presence of all-trans retinal, a retinal isomer ubiquitously present in every tissue, like microbial rhodopsins and unlike canonical vertebrate opsins. Remarkably, transgenic worms expressing MosOpn3 were ~7,000 times more sensitive to light than transgenic worms expressing ChR2 in this light-induced behavior, demonstrating the advantage of MosOpn3 as a light switch. LamPP is a UV-sensitive bistable opsin having complete photoregenerative ability by green light. Accordingly, transgenic worms expressing LamPP in cholinergic motor neurons stopped moving upon violet light illumination and restored coordinate movement upon green light illumination, demonstrating color-dependent control of behavior using LamPP. Furthermore, we applied molecular engineering to produce MosOpn3-based tools enabling light-dependent upregulation of cAMP or Ca2+ levels and LamPP-based tool enabling clamping cAMP levels color dependently and context independently, extending their usability. These findings define the capacity of two bistable opsins with similar retinal requirement as ChR2, providing numerous strategies for optical control of various GPCR-based physiologies as well as GPCR signaling itself.

Cucumber powdery mildew detection method based on hyperspectra-terahertz.

To explore the use of information technology in detecting crop diseases, a method based on hyperspectra-terahertz for detecting cucumber powdery mildew is proposed. Specifically, a method of effective hyperspectrum establishment, a method of spectral preprocessing, a method of selecting the feature wavelength, and a method of establishing discriminant models are studied. Firstly, the effective spectral information under visible light and near infrared is preprocessed by Savitzky-Golay (SG) smoothing, discrete wavelet transform, and move sliding window, which determine the optimal preprocessing method to be wavelet transform. Then stepwise discriminant analysis is used to select the feature wavelengths in the visible and near-infrared bands, forming the feature space. According to the features, a linear discriminant model is established for the wave bands, and the average recognition rate of cucumber powdery mildew is 93% in the whole wave band. The preprocessing method of terahertz data, the screening method of terahertz effective spectrum, the selection method of feature wavelength and the establishment method of classification model are studied. Python 3.8 is used to preprocess the terahertz raw data and establish the terahertz effective spectral data set for subsequent processing. Through iterative variable subset optimization - iterative retaining informative variables (IVSO-IRIV), the terahertz effective spectrum is screened twice to form the terahertz feature space. After that, the optimal regularization parameter and regularization solution methods are selected, and a sparse representation classification model is established. The accuracy of cucumber powdery mildew identification under the terahertz scale is 87.78%. The extraction and analysis methods of terahertz and hyperspectral feature images are studied, and more details of lesion samples are restored. Hence, the use of hyperspectral and terahertz technology can realize the detection of cucumber powdery mildew, which provides a basis for research on the hyperspectral and terahertz technology in detection of crop diseases.

Functional identification of an opsin kinase underlying inactivation of the pineal bistable opsin parapinopsin in zebrafish.

In the pineal organ of zebrafish larvae, the bistable opsin parapinopsin alone generates color opponency between UV and visible light. Our previous study suggested that dark inactivation of the parapinopsin photoproduct, which activates G-proteins, is important for the regulation of the amount of the photoproduct. In turn, the photoproduct is responsible for visible light sensitivity in color opponency. Here, we found that an opsin kinase or a G-protein-coupled receptor kinase (GRK) is involved in inactivation of the active photoproduct of parapinopsin in the pineal photoreceptor cells of zebrafish larvae. We investigated inactivation of the photoproduct in the parapinopsin cells of various knockdown larvae by measuring the light responses of the cells using calcium imaging. We found that GRK7a knockdown slowed recovery of the response of parapinopsin photoreceptor cells, whereas GRK1b knockdown or GRK7b knockdown did not have a remarkable effect, suggesting that GRK7a, a cone-type GRK, is mainly responsible for inactivation of the parapinopsin photoproduct in zebrafish larvae. We also observed a similar knockdown effect on the response of the parapinopsin photoreceptor cells of mutant larvae expressing the opsin SWS1, a UV-sensitive cone opsin, instead of parapinopsin, suggesting that the parapinopsin photoproduct was inactivated in a way similar to that described for cone opsins. We confirmed the immunohistochemical distribution of GRK7a in parapinopsin photoreceptor cells by comparing the immunoreactivity to GRK7 in GRK7a-knockdown and control larvae. These findings suggest that in pineal photoreceptor cells, the cone opsin kinase GRK7a contributes greatly to the inactivation of parapinopsin, which underlies pineal color opponency.

Color opponency with a single kind of bistable opsin in the zebrafish pineal organ.

Lower vertebrate pineal organs discriminate UV and visible light. Such color discrimination is typically considered to arise from antagonism between two or more spectrally distinct opsins, as, e.g., human cone-based color vision relies on antagonistic relationships between signals produced by red-, green-, and blue-cone opsins. Photosensitive pineal organs contain a bistable opsin (parapinopsin) that forms a signaling-active photoproduct upon UV exposure that may itself be returned to the signaling-inactive "dark" state by longer-wavelength light. Here we show the spectrally distinct parapinopsin states (with antagonistic impacts on signaling) allow this opsin alone to provide the color sensitivity of this organ. By using calcium imaging, we show that single zebrafish pineal photoreceptors held under a background light show responses of opposite signs to UV and visible light. Both such responses are deficient in zebrafish lacking parapinopsin. Expressing a UV-sensitive cone opsin in place of parapinopsin recovers UV responses but not color opponency. Changes in the spectral composition of white light toward enhanced UV or visible wavelengths respectively increased vs. decreased calcium signal in parapinopsin-sufficient but not parapinopsin-deficient photoreceptors. These data reveal color opponency from a single kind of bistable opsin establishing an equilibrium-like mixture of the two states with different signaling abilities whose fractional concentrations are defined by the spectral composition of incident light. As vertebrate visual color opsins evolved from a bistable opsin, these findings suggest that color opponency involving a single kind of bistable opsin might have been a prototype of vertebrate color opponency.

[Research on distinguishing weed from crop using spectrum analysis technology].

Automatic detection of weeds is necessary for site--specific application of herbicides or precise physical weed control. Leaf reflectance is mainly determined by photosynthetic pigments, leaf structural properties and water content, so spectral reflectance characteristics can be used for weed discrimination. The spectral reflectance of cotton, rice and weeds was determined in the range from 350 to 2 500 nm using the Analytical Spectral Device Full Range FieldSpec Pro (ASD) in laboratory. The discrimination analysis was done using the statistical software package SAS. The characteristic wavelengths were selected by using STEPDISC procedure. With the selected characteristic wavelengths, discriminant models were developed using the DISCRIM procedure in SAS. For distinguishing spine-greens from cotton, three characteristic wavelengths, 385, 415, and 435 nm, were selected, and good classification performance (100% accuracy) was achieved. The combination of characteristic wavelengths 415 and 435 nm has the biggest contribution to discrimination model. For distinguishing barnyard-grass from rice, five characteristic wavelengths, 375, 465, 585, 705, and 1 035 nm, were selected, and also good classification performance (100% accuracy) was obtained. The transition point from yellow to orange wavelength (585 nm) and the wavelength 705 nm in the red edge contributed more to discrimination model.